Everyone listed in an Animal Use Protocol form must know the material covered in this section and successfully pass the test. If someone has had training in another institution or has extensive experience and believes that they are familiar with the information, asking for the test and passing is sufficient to obtain certification.
Content of the Basic Level:
- Biological hazards
- Physical injury
- Occupational Health Program (OHS)
- Personal hygiene and protection
- Factors influencing research results
- Stress reduction
- Environmental enrichment
- Controlled substances
- Other issues
Regulations, Animal Welfare and Protocol Approval Process
Our institution follows requirements from federal, provincial and municipal legislation. This would include, for example, the federal Health of Animals Act (38-39 Elizabeth II, Chapter 21, pgs. 387-421, laws.justice.gc.ca/en/H-3.3/ ), which replaced the Animal Disease and Protection Act and which governs control of animal diseases and toxic substances. Those working with animals should be cognizant of institutional and/or facility safety program and are strongly recommended to enroll in the McGill Occupational Health Program for Animal Related Activities (see section specific to this program).
Workplace Hazardous Materials Information System (WHMIS), which resulted from federal and provincial co-operation, was instituted in 1988. Federal government laboratories are governed by federal WHMIS and the Canada Labour Code. The following publications are available free of charge from Labour Canada: The Employer and WHMIS; Introduction to the WHMIS Program; Exercise WHMIS in the Workplace, and a relevant poster. McGill and affiliated hospital give regular WHMIS sessions. Contact the McGill Environmental Health and Safety Office’s Web site.
Elsewhere, provincially enacted Health and Safety legislation specifies the accountability of directors and the rights and responsibilities of employers, supervisors and workers in the workplace. The right to refuse unsafe work is a part of the Occupational Health and Safety (OHS) Act (see CSST Web site). WHMIS regulations are also a section of this legislation and require that each employer provide safe working conditions and that employees be informed about all hazards they will face in the course of their duties. Employees are also given the right to withdraw from the workplace if faced with an unsafe condition. All hazardous substances, including microorganisms, must be labeled in a specified manner, and a Material Safety Data Sheet (MSDS) must be available to accompany each hazardous substance. Each province has adapted these federal government guidelines for its own purposes. WHMIS material may be obtained from the McGill Environmental Health and Safety Office.
MCGILL AND AFFILIATED HOSPITAL ANIMAL USE APPROVAL PROCESS:
Submission of planned research or teaching project
If the category of invasiveness is "A", no approval is needed. "A" level includes studies or experiments on most invertebrates or no entire living material (such as insects, eggs, animal tissue or skin). Also "A" level is no-contact studies such as bird watching.
All "B", "C" and "D" category of invasiveness using vertebrates and higher cephalopods require approval prior to starting the project.
To obtain approval, the Animal Use Protocol form must be completed and approved by the Facility Animal Care Committee (FACC) via the on-line Darwin software. If you do not have access yet, please submit a request via this Webform.
The submitted protocol, or amendment, is reviewed by members of the FACC. This committee ensures that the institution's as well as the CCAC guidelines and policies are followed. The membership includes: at least two faculty members, a veterinarian, a community representative, an institutional member not using or depending on animals, a technical staff representative, research ethics officer and a graduate student.
The protocol will either be:
- approved as is,
- the committee makes recommendations for specific changes before approval is given,
- it is not approved
Know that once approved, a project can still be halted under certain conditions. If an animal is found to be experiencing unrelenting pain and distress, the FACC and Veterinarian have the authority to have the animals euthanized and/or terminate the procedures.
All projects must show proof of potential benefit for humans and/or animals to justify using living animals. This is achieved when a recognized peer-reviewed agency (such as CIHR and NSERC) awards a grant for the project. Applications for grants are very competitive and only the very best get funded. Note: the agency's name and grant title must appear in the funding section of the animal use protocol.
If the project is funded by a non-peer-reviewed source OR if the investigator would like to start the project before the funding by a peer-reviewed agency is awarded, the researcher can request internal peer review from the Associate Dean of his/her Faculty or from the Associate Director of the Research Institute of his/her Affiliated Hospital.
Standard Operating Procedures (SOP)
SOP stands for Standard Operating Procedures. There are procedures that are used often and the details have been written down in order to help investigators with the details concerning each of them. These SOPs are already approved by the institution and the CCAC and this helps in maintaining a high standard and speed up protocol review. Another important point is that each procedure and animal is treated the same way.
The SOPs are downloadable from the UACC Web site (www.mcgill.ca/research/researchers/compliance/animal/sop ).
Length of time for the entire approval process
The entire process should take about a month in order for the Facility Animal Care Committee to review and meet, corrections be made by the investigator if necessary and final processing be done.
Approval is valid for 1 year.
Principal Investigator's responsibility
On the Animal Use Protocol form, the principal investigator must agree to the statement:
"“The information in this application is exact and complete. I declare that all care and use of animals in this proposal will be in accordance with the guidelines and policies of the Canadian Council on Animal Care and those of McGill University. I shall request the Animal Care Committee’s approval prior to any deviations from this protocol as approved. I understand that this approval is valid for one year and must be renewed on an annual basis. Approval from the committee does not guarantee availability of space, equipment and services; contact the animal facility supervisor for requirement. Everyone listed in the protocol must have read the Animal Use Protocol's sections pertinent to what they are mandated to perform.”"
Everyone listed in the personnel section for the project must have read the approved protocol and respect the above mentioned agreement.
Common errors in completing the form
The items that present the most problems when a protocol gets reviewed by the Facility Animal Care Committees are:
- Section “5- Keywords of Procedures” = keywords only, not sentences. Why? A report is sent every year to the CCAC and this is the format they have requested.
- Section “19- Justification For Number Requested” = total numbers of animals stated in the table must be the same as the totals described in the justification of animal numbers section. Basically, what is needed for the animal number justification, is the simple arithmetic on how the totals in the table were arrived at.
- Section “31- Adverse Effects and Humane Intervention Points” = As known as “Clinical endpoint”: can be defined as the point at which an experimental animal's pain and/or distress is terminated, minimized or reduced by taking actions such as humanely killing the animal, terminating a painful procedure, or giving treatment or relieve pain and/or distress" The importance of clear and measurable criteria for the clinical endpoint is because the animal user must know exactly at what point, the animal's distress has become unacceptable and action must be taken to end its suffering. Specific clinical criteria is needed, statements such as "if the animals is sick..." or "the technician takes care of the animal's health" are not acceptable. Good criteria: % of weight loss, maximum tumour size, absence of grooming and anything that can be expected when doing the experiments.
- Section “32- Description of Procedures” = It is important to include experimental endpoints which is defined as the estimated survival time for the animals.
- Section “29- Euthanasia" = if using a physical method of euthanasia without the use of anesthesia, it must be justified.
Basic Animal Care
A. GENERAL PRACTICES
The portal through which animals and containers enter a facility is important in the overall prevention of disease. The examination of newly arrived animals should have several aims: evaluating the condition and health of the animals; preventing cross-contamination of animals from different sources; and ensuring that the order has been accurately filled. The health status of the animals at their source and the possibility of cross-contamination during transport are important considerations. Cross-contamination is always a greater risk if the animals are not shipped in a vehicle dedicated to the transport of animals, coming from a single source. However, this risk can be decreased by the use of filtered shipping crates.
Each new shipment of animals should be received, uncrated and examined by trained personnel and placed in clean cages in a designated reception area separate from the animal holding room(s). The reception area should be cleaned and disinfected after each new shipment. Shipping containers should not enter the main facility unless properly decontaminated and should be properly disposed of, or thoroughly cleaned and disinfected if they are to be reused. Incoming animals should be identified and their arrival appropriately recorded.
Animals that appear unhealthy, or which have been in any way debilitated in transit, should be separated from the remainder and held in an appropriate location for observation and treatment. Where this is not feasible, such animals should be euthanized without delay.
Cell lines should be shown to be free of specific organisms before they are injected into disease-free animals.
Quarantining is most important as it allows detection of disease prior to introduction to a disease-free colony. The level of conditioning required will depend on the differences in microbial status between the resident animals and the incoming animals. Rodents are commonly purposebred for the laboratory and can be received from the supplier in a defined state, with a known health, nutritional and, to a varying extent, genetic background. Similarly other purposebred species obtained from reputable sources will have complete health profiles and will have received specified prophylactic treatment. Such animals may not normally require further quarantine for confirmation of their health status; however, a holding period of several days will give the animals the opportunity to adjust to their new surroundings. A minimum adjustment period of two days is required after shipping for immune function, corticosterone levels and other physiological parameters to stabilize.
Legally procured strays or donated animals, acquired feral animals, such as non human primates, and animals from other random sources should be subjected to a period of conditioning following their reception. Conditioning requires that the animal be held for a varying period of time (1-6 weeks) in separate quarters. The length of the conditioning period will depend on the species, the health status of the animals, the reliability of the supplier, and whether an active process of screening animals for the presence of or exposure to infectious agents is undertaken. During this period, thorough physical examinations should be undertaken. Further examination will depend on species and intended use.
The conditioning period should be of sufficient duration to permit the proper evaluation of the suitability of the animal(s) for their intended use, and testing for contagious, zoonotic and other diseases that may be of concern. This can include serological screening for antibodies to viruses and other pathogens, the examination for ecto- and endo-parasites, mycoplasma, and pathogenic bacteria. If contamination during transport is a possibility, an appropriate period to allow expression of disease or antibody production should be allowed. Sufficient time must also be allowed for appropriate treatment or vaccination against diseases that tend to be endemic in the species being conditioned.
During the conditioning/quarantine period, the animals should be held in facilities separated from other animals with no crossover of personnel, equipment, supplies or ventilation, unless effective measures are taken to prevent cross-contamination. Wearing facility-specific protective clothing helps keeping animals free of infectious disease.
3. Holding (Maintenance)
Only one species should be housed in a conventional animal room unless maintained in isolator caging, racks or cabinets. Shipments of the same species, acquired from different suppliers, should also be separated according to health status if space permits, or housed in isolator caging. Where the mixing of species and/or stocks from different sources is unavoidable, every effort should be made to place together those that are behaviourally compatible, have similar environmental requirements, and a low probability of cross-infection. Non human primates should not be housed with any other species.
HEPA (High Efficiency Particulate Air) filters are often used in animal facilities to filter out all the particles in the air down to tenths of a micron in diameter.
Animals should be held in enclosures adequate for that species as described in the CCAC’s Guide to the Care And Use of Experimental Animals, Volume 2 (http://www.ccac.ca/en_/standards/guidelines/additional/vol2_mice).
4. Identification and Records
Cage or group identification may be used for small laboratory animals if individual identification is not an experimental prerequisite. Individual identification can be by ear tagging, ear notching, tattooing, tail marking, subcutaneous microchip implant or other species appropriate method. Dye marking on the hair provides for short-term identification. Larger laboratory animals should always be individually identified by tattoo, neck band, individual tag, or a subcutaneous identity tag.
The Canadian Council on Animal Care (CCAC, www.ccac.ca) opposes the use of toe clipping as a method of identification for the short-term learning experience of field studies. Where it is necessary to provide permanent individual identification between litter members of newborn rodents, toe clipping may be necessary. If, for any reason, this procedure has to be undertaken on other than neonatal animals, either a local or general anesthetic should be administered.
The importance of keeping complete and thorough records on all experimental animals cannot be overemphasized. Monitoring of the animal's health and of the experiment's progress usually depends on knowing which animal is which. The following information should be recorded for each animal: arrival date, sex, estimated age and weight, breed and type, colour and markings and any physical abnormalities or other identifying features. The name of the project or investigator and protocol number to which it is allocated should be noted, as well as that of the supplier and eventual disposition. Animal records should be kept for a period of one year after the final disposition of the animal. The cages in which animals are housed, prior to or during an experiment, should be clearly marked indicating the sex and number of contained animals, the investigator responsible for them and such special instructions as may be pertinent to their care. Records, especially when used in conjunction with data processing equipment, can facilitate facility management.
Use of room cards/boards on the doors of animal rooms indicating the species, investigator(s) whose animals are being held and any special notations that may be pertinent is a good practice.
People donating animals to research facilities are required to sign a statement that they are the legal owner. This document should include identification of the animal by the criteria previously noted, and should specifically transfer ownership and disposal of the animal to the institution. Animals (such as dogs, for which a system of national registry exists) should always be checked for the presence of identifying markings.
B. CARE OF THE ANIMAL
Every species has its own need regarding its environment, food requirement and correct handling by the animal user. It will be distressed if it is not provided with the minimum it needs for living. Therefore, for ethical reasons as well as to ensure that stress will not affect the results of the study, animal husbandry is important.
All experimental animals should receive palatable, wholesome and nutritionally adequate food according to the requirements of the species, unless the study requires otherwise. In certain experiments where small quantities of chemical residues may influence results, certified diets with documented analysis of contaminant pesticides, herbicides, etc., are available from commercial manufacturers of laboratory animal diets.
a) Food Storage
Whenever possible, pasteurized or sterilized diets obtained from reputable suppliers should be used. Proper storage of foods is necessary to minimize the possibility of contamination, deterioration or spoilage. Dry laboratory animal diets should be used within six months of the milling date when stored in cool, dry, well-ventilated quarters. Irradiated diets kept under the same conditions have approximately double the shelf life. Primate and guinea pig diets should be used within three months of the milling date, unless vitamin C is supplemented. To avoid problems from age deterioration, the date of milling of each shipment should be obtained from the supplier (this is usually marked in code on the bags). Bags should then be marked, put on plastic or metal pallets or racks to keep them off the floor, and stored so that the oldest will be used first. Stale shipments should not be accepted. Shelf life will be appreciably enhanced if the storage area is maintained at a temperature of <16C (60.8F). Canned foods can be safely stored for long periods. Clean green vegetables suitable for human consumption may enhance the diet; however, vegetable discards may prove sources of infection and should be avoided.
Food used in microbe-controlled environments is often autoclaved. Autoclaving decreases the concentrations of some vitamins and antioxidants. However, autoclavable diets are available which contain higher concentrations of heat-labile ingredients to compensate for the losses induced by heat-sterilization. Shelf life may be decreased, but need not be if the process is handled properly. Gamma irradiation is also used for diet sterilization.
Diet in large quantities should not be stored in animal holding rooms. Small quantities, sufficient for one or two days may be kept in the room in covered, vermin-proof containers.
b) Special Considerations
All animals tend to reduce their food intake when sick. Animals with a high metabolic rate, e.g., small rodents and those requiring fairly frequent feedings of high protein diets (e.g., the cat), can become debilitated very rapidly. In cases of anorexia in these species, oral intubation and force feeding as well as intravenous therapy (cat) should be instituted without undue delay. Restricted feeding for maintenance of adult animals is commonly practiced for some species and strains, such as rabbits. Animals on restricted food or fluid intake for experimental purposes should be closely monitored for weight loss, signs of dehydration, signs of stress and deterioration in health. It should be noted that food and water restriction may have a marked effect on the response of animals to toxic substances and other experimental variables. For some species, particularly non human primates, providing a variety of foods can be useful as a form of environmental enrichment.
Generally, food should not be scattered over the bottom of the cage, where it may be contaminated or wasted. Exceptions to this include provision of food to newly hatched birds and abnormal (handicapped) animals, such as mice with muscular dystrophy.
Drinking water should be available to animals at all times, unless contra-indicated by the experimental protocol. Tap water, even if from municipal water systems, is not sterile and quickly becomes contaminated with even more bacteria after the bottle is placed on the cage. Monitoring water quality is an important aspect of any research program, as water contamination and chemical composition can affect the health of animals and the results of animal experiments.
Methods available to remove both microbial and chemical contamination include acidification, chlorination, reverse osmosis, ultrafiltration and ultraviolet (UV) light. Some of these methods may alter immune function and growth rates in experimental animals. Regardless of whether or not the water supply is treated, all water dispensing equipment should be thoroughly sanitized according to institutional SOPs, and periodically monitored for bacterial contaminants.
A watering method unlikely to spread disease or contaminate the water supply should be chosen. Water bottles should be transparent so as to permit ready observation of cleanliness and water level; of a material that will withstand sterilization, and of a wide mouth design to facilitate cleaning. Water bottles should always be replaced with clean, freshly filled ones, rather than by refilling the ones in use. Animals housed under freezing conditions may require heated water bowls.
Automatic watering devices are economical to operate, but if not properly designed, are difficult to disinfect properly and may lead to cross-contamination. Recirculating systems eliminate stagnation of water and help prevent buildup of microorganisms. The correct pressure in the drinking valves prevents backflow of water into the lines when animals drink from or play with the valve. Malfunction of automatic watering systems can lead to drowning or drought; consequently, the system must be routinely and thoroughly checked. Some animals need to be taught to use automatic watering devices. Automatic watering devices are not recommended for guinea pigs, unless they are habituated to them.
Most fish have a low tolerance for both copper ions and chlorine. Their water supply, therefore, should either be dechlorinated or obtained from an untreated source, and should not be brought into the aquarium through copper piping.
Experts disagree about the need for exercise in laboratory animals. A judgment in such cases must thus be made by the laboratory animal veterinarian in consultation with the investigator. Although many adult animals do not seem to have a motivation to exercise per se, in the process of satisfying their behavioural needs, they do get exercise. Exercise requirements for animals should reflect species, age and environment. Research information on the requirements of each species for exercise is limited and varied, but continually increasing. Young animals of most species involve themselves in much more play and exercise activity than adults. For some species, exercise may not be required in adult animals for physiological health. Several studies suggest that there are no beneficial effects on behaviour, health or in enhancement of voluntary activity in the laboratory-bred beagle from increasing the cage dimensions beyond the standard 76 cm x 76 cm x 76 cm (30" x 30" x 30") size, provision of half-hour daily exercise, or from 1.22 m x 3.05 m (4' x 10') floor pen housing. Judgment should be based on the animal's breed, temperament, physical condition, the conditions under which it has previously been kept and the length of time it is to be confined. Animal cages must, however, always be large enough to allow the innate normal behavioural and postural adjustments. There are many varied methods and programs of exercise which are successfully used in dogs, including walking programs using outside volunteers. Caged rats spontaneously exercise by playing with cage mates and during feeding.
C. CARE OF THE FACILITY
1. Cleaning and Sanitation
Employees must be aware of proper cleaning and disinfecting procedures and their importance in disease prevention. All cages, pens, racks, aquaria, accessory equipment, etc., must be thoroughly cleaned and disinfected before reuse. Most of these items should be subject to regular (usually weekly) cleaning during use. As a general rule, laboratory animals should be moved to freshly cleaned cages at least once a week. Cleaning practices need to be modified according to the species and housing system for domestic animals, fowl, reptiles and aquatic animals. The effectiveness of detergents, disinfectants and facility cleaning programs should be monitored and constant.
The ability to clean and sanitize a facility is greatly influenced by facility design and construction materials. The objective of a sanitation program is to reduce the microbial contamination or "bioburden" to a level that reduces the possibility of any cross-contamination. Proper sanitation will not compensate for the transfer of infection by personnel. Cleaning and sanitation merely complement proper procedures which minimize contamination. Activities such as pressure spraying and dumping bedding can aerosolize microorganisms allowing cross-contamination if animals are present. Opening doors can alter the airflow in a facility, enhancing the possibility of transfer of contaminants. Moveable equipment can transmit organisms between areas. Therefore, such equipment should be dedicated to a particular room or area.
Procedure rooms using animals from different sources are a potential source of cross-contamination. Proper disinfection of surfaces should be ensured after use.
Bedding in animal cages or pens should be changed as often as necessary to keep the animal clean, dry, and relatively odour-free and ammonia levels in the cage at appropriate levels. In rats, this is 25 ppm. Smaller laboratory animals require one to three changes per week, depending on such variables as the sizes of the animals, population density and type of caging and whether or not litters are being produced. Larger species such as dogs, cats and non human primates usually require at least a daily change.
Food containers should be easily cleaned and disinfected.
Animal cages are most efficiently cleaned and sanitized with mechanical washing equipment operating at 83C (180F) or higher, for a minimum of ten minutes. Cages should be carefully rinsed to remove all traces of washing and disinfecting agents, as exposure to these may adversely affect both the animal and the experimental results. All automatic washing equipment should be subjected to regular maintenance to assure proper performance. Where an automatic cagewasher is not available, use of a spray washer and disinfectant are preferable to the dip tank and rinsing method. It should be noted that sodium hypochlorite and iodophores are effective on most animal viruses; however, disinfectants should be chosen according to the spectrum of viruses and organisms required to be killed and the possibility of deactivation by the local environment. There are references available to aid in identification of the appropriate disinfectants. Chlorine dioxide sterilants/disinfectants have become more recently available and are often used in facilities maintaining SPF or immunosuppressed animals because of their rapid broad spectrum activity, even in the presence of an organic load.
All chemicals should be used properly, according to label directions. Detergents, disinfectants and pesticides may cause changes in the experimental animal by inducing or inhibiting cellular enzyme activity. This should be a consideration when conducting experiments which may be adversely affected.
2. Waste Disposal
Dead animals, animal tissues and excreta, bedding, unused food, etc., should be collected in leak-proof metal or plastic containers with leak-proof, disposable liners and tight lids. Liners are essential for animal tissues, carcasses, and radioactive or toxic waste. Infectious waste should, ideally, be incinerated on the site. If the waste is to leave the facility it should be sterilized (autoclaved) before removal. Gamma irradiation is a relatively recent method of disinfection of waste products which may come into more prominent use.
Waste which cannot be rapidly disposed of should be stored in a cold storage area provided for that purpose. Such areas must be vermin-free, easily cleaned and disinfected as well as being physically separated from other storage facilities. The waste storage area should be located so that wastes need not be carried through other rooms of the facility.
Dead animals should be removed from cages as soon as they are noticed. The laboratory animal veterinarian who should have been immediately informed of sick animals, should also be informed of dead ones. Dead animals should be properly identified, placed in disposable plastic bags and taken to the postmortem area immediately upon discovery. In the postmortem area they should be held under refrigeration for necropsy or for disposal in accordance with the investigator's instructions. National guidelines as well as local and provincial laws control waste disposal practices that could endanger public health.
Considerable forethought and extensive consultation is advisable before installing an incineration facility for the disposal of pathological waste.
3. Vermin Control
A properly constructed building should be vermin-proof, but may not be free from vermin. Vermin enter on food, bedding, people and animals. Insects and arthropods thus introduced, may act as the intermediate hosts of certain parasites and may also mechanically transmit bacterial and other pathogens. Wild rodents may transmit a wide variety of bacteria, viruses, and parasites to caged members of closely related species. New facilities should be checked critically for vermin before any animals are moved in.
Vermin should also be controlled in already-infested older buildings. A control program will include the proper training of personnel, good waste disposal, sealing or eliminating breeding sites, extermination through pesticides or trapping, and the recovery of all escaped and/or wild animals. It is important that pesticides be applied only under professional supervision. Many pesticides are dangerous to humans, and may adversely affect the experimental animal and the research. Any control program that is initiated must extend throughout all areas of the facility, with special attention to food and bedding storage. The practice of using a free-roaming cat for the control of wild and escaped rodents is not acceptable except in farm animal facilities, and only under close management.
If insect colonies are kept in or near an animal care facility, there must be regular monitoring of the facility against infestation from escapees. Such insect colonies should be kept behind a screened enclosure or inside an escape-proof container. The use of insecticides must also be compatible with these insect colonies.
4. Holiday and Emergency Care
- Weekend and Holiday Care of Laboratory Animals is Essential. It should be recognized that changes in personnel and feeding and cleaning schedules, as can occur during these periods, are known to be stressful to routine-oriented animals.
- Animal Care is a Continuous and Daily Responsibility
This point should be emphasized in job descriptions for animal care personnel and in union contracts. Basic animal care should be categorized as an "essential service" and a clause to this effect should be included in all collective agreements, and should not be subject to interruption through strike action. Staff must be provided for weekends and holidays, and skilled assistance must be available in the event of an emergency.
The names and telephone numbers of staff responsible for the animals should be given to security personnel. Some institutions may also choose to have contact telephone numbers posted prominently in the facility. In either case, directions for contacting responsible animal care staff must be made available in the facility. All the animal care staff should be informed of their responsibilities in emergency situations.
Where employees are unionized, the CCAC suggests an essential services provision to be inserted near the strikes and lockouts clause for the designation of employees to care* for research animals. The parties must agree that proper care of all research animals will be maintained by the members of the bargaining unit in the event of a strike or lockout in the course of the agreement or its continuance.
At least seven days before the commencement of a strike or lockout, the employer will designate and identify a number of employees which it deems sufficient to provide for continuous proper care of the animals during the strike or lockout. A list of the names must be delivered to the Union and the parties agree to meet with a view to executing a formal agreement with respect to the employees affected. Should the parties be unable to reach agreement on the persons to be designated, the matter will be referred to the CCAC, for final and binding resolution by the Council.
All persons so designated must be paid their regular salary during the period of designation.
Due regard should be had for previously arranged vacations and other matters and as far as possible the designated duties will be dispersed among all appropriate employees equally. No other duties will be assigned to these designated employees.
* Proper care implies provision of appropriate temperatures, humidity, light cycles, ventilation, food, water and cleaning as well as exercise and nursing care where appropriate
Ethics of Animals in Research
Why animals play a crucial role in research
From the development of insulin to the latest life-prolonging cancer drugs and virtually every major medical advance in between, animals have played vital roles in scientific research that have led to cures and treatments for a wide array of human diseases. They have helped scientists improve the nutritional value of our food supply and – thanks to agricultural and veterinary research – have helped bring about a better quality of life for many animals and a safer environment as well.
Millions of lives have been saved, improved and extended thanks to the results of humane scientific research that has relied upon animals at various stages. Without the use of animals, men, women and children around the world would simply not enjoy the quality and length of life they do today.
The “Three Rs”
Animals are used in research when there is simply no alternative that will produce the necessary results. But before scientists at McGill are allowed to employ animals in research, they must follow what are called the “Three Rs” as established by the federal government’s Canadian Council on Animal Care (CCAC). That means:
- · they must replace animals with alternative research methods wherever possible
- · they must reduce the use of animals to the least number possible
- · they must refine their procedures to minimize adverse conditions for animals
It isn’t easy to get a research proposal involving animals approved at McGill. Before animals are involved in research, two levels of review must occur: First, a peer research panel must determine that the proposed research project does indeed have scientific merit and that it can lead to advances in understanding and knowledge. Second, the University’s Animal Care Committee must approve the project. Even after independent research experts and the Animal Care Committee approve, the researchers must then follow strict guidelines imposed by the Canadian Council on Animal Care. At McGill, we meet or exceed federal guidelines and our facilities are inspected regularly.
Here’s how the process works:
MULTI-STEP PROCESS ENSURES THAT RESEARCH WITH ANIMALS IS VERY CAREFULLY MONITORED AT McGILL UNIVERSITY
The federal government’s Canadian Council on Animal Care oversees every aspect of research involving animals. It inspects all animal facilities, reviews the work of university committees, reviews research projects and reviews institutional policies every three years
1. Researcher submits a scientific project proposal to an agency. The proposal gets evaluated and only the most meritorious will be supported and the use of animals will only be considered if it is established that there is no other way of achieving the research objectives.
2. If the scientific project proposal was found to be meritorious, the researcher submits a proposal to the institution to do animal research before starting a new project or renewing an existing one
3. The University’s Animal Care Committee (at a minimum composed of a veterinarian, researchers, a community representative, animal care staff and a compliance officer) reviews all aspects of the project with emphasis on ensuring animals will receive the best care possible for achieving the research objectives. It rejects, approves or gives conditional approval to the procedures in the project or to changes in the procedures before allowing the research to proceed. This review is about the welfare of the animals.
4. Mandatory training of personnel on animal handling and procedures as well as health precautions are assured for all research personnel and animals
5. The institution approves the project for one year
6. Assistance is available to refine procedures, train people, care for the animals and, when needed, make changes to the research project
7. Quality Assistants ensure that research personnel follow the approved proposal
8. One year later, the researcher must submit another proposal
in order to continue the research
Researchers and everyone involved in research with animals – including veterinarians and animal-care technicians – are sincerely concerned about the welfare of animals that are part of the research process. But researchers are also concerned about the sick and disabled among us who are desperate for ways to deal with pain or the prognosis of fatal illness or who seek better ways to ease their suffering from a chronic medical condition.
Thousands, perhaps millions, of lives can be improved by a successful research project that leads to better care and treatment – for the grandfather taken by Alzheimer’s disease, the mother stricken with breast cancer, the child learning to live with diabetes, the whole segment of a community trying to cope with excessive levels of cholesterol or heart disease. Those are the people the researchers are trying to help.
Myths and realities
A number of myths or misconceptions have arisen in the discussion about the involvement of animals in scientific research. It is important to know the facts.
Myth: Animals are not needed in research.
Fact: Whereas every effort is made to minimize the use of animals, at certain stages of research projects, a living organism must be tested before a drug or treatment is approved for human trials. Most people would consider allowing human trials of new drugs or procedures without prior testing on animals to be dangerous and unacceptable.
Myth: Studying animals does not provide insight into human health
Fact: Genetic and physiological similarities between humans and animals provide researchers with irreplaceable and invaluable insights into how human systems might react to a drug or treatment.
Myth: Dogs, cats and monkeys are the most widely used animals in research.
Fact: Fish and rodents, usually mice or rats, account for more than 83% per cent of the animals used in research and are bred specifically for research purposes. Stolen pets or SPCA animals (other myths) are not used in research. Dogs and cats are purchased from reputable suppliers.
Myth: Research animals live in near-constant pain and suffering
Fact: The vast majority of biomedical research does not result in significant discomfort or distress for research animals. The 2008 report of the Canadian Council on animal care shows that the overwhelming majority of procedures involving animals are described as experiments that cause little or no discomfort or stress or experiments that cause minor stress or pain of short duration such as an injection or minor surgery similar to pets undergoing spay or neutering.
How animals have helped
At McGill, recent research involving animals has led to the following advancements:
- · the discovery of previously unknown interactions between genes that control whether cells become cancerous
- · the development of new, experimental treatments for diseases that affect the nervous system
- · a better understanding of the mechanisms of blood flow that has helped in the development of drugs to ease vascular head pain
Other familiar breakthroughs involving animal research have included:
- · the discovery of insulin, penicillin, streptomycin and yellow fever vaccine
- · the treatment for cancer, aids, hypertension, cardiac stents, high cholesterol, depression
- · the development of such important medical devices as the electrocardiogram, computer assisted tomography (CAT scan) and magnetic resonance imaging (MRI)
- · improved understanding of how cells work, how genetic differences play a role in the development of life and disease, immunity and the regulation of cholesterol
There are, in fact, too many medical research breakthroughs to list them all here. But it is accurate to say that cancer patients are living longer, HIV sufferers are living longer, diabetics are living longer thanks to research that has involved animals. Those diseases used to be automatic – and almost always rapid – death sentences. Now, they needn’t be.
Animals in laboratories around the world have played an invaluable role in helping us understand disease and what we need to do to treat it or cure it.
Where to find out more
There is a wealth of information about humane animal research available on the Internet. Here is a sample of sites you might want to visit to learn more:
McGill University and Affiliated Hospitals’ animal care program: www.mcgill.ca/research/researchers/compliance/animal
Canadian Council on Animal Care (federal government): www.ccac.ca
Canadians for Health Research: www.chrcrm.org
Foundation for Biomedical Research: www.fbresearch.org
Student website on research: www.speakingofresearch.org
Another student-supported site on the benefits of research: www.pro-test.org.uk
The Society for Neuroscience: www.sfn.org
You can also send an animalcare [at] mcgill [dot] ca (email )if you have more questions or comments.
Occupational Health & Safety
Projects often include the use of hazardous material. Such agents as carcinogens, transplantable tumours, radioactive isotopes, toxic chemicals and infectious material require that warnings and precautions to protect both animals and personnel be included in the project and described in detail in the biohazard section of the animal use protocol. The investigator must find out about the potential hazard of the agent he/she will be using and the ways to protect the animal users, clean the equipment and spills and how to handle animal waste.
Guidelines for working with biohazards (e.g., bacteria, viruses, parasites, fungi and other infectious agents), are provided in the Public Health Agency of Canada 's Laboratory Biosafety Guidelines. The guidelines include such items as biohazard containment, laboratory design, personal hygiene and safety facilities, and can be used to provide training for employees as mandated by WHMIS (Workplace Hazardous Material Information System, http://www.hc-sc.gc.ca/ewh-semt/occup-travail/whmis-simdut/index-eng.php ).
The biosafety guidelines apply to all research carried out or supported by the federal government and have been adopted by many industries.
Standard Operating Procedures (SOP) based on the guidelines, aimed at minimizing risks to humans in biohazard risk areas, have been developed and are enforced. The SOPs can be found on the UACC Web site at www.mcgill.ca/research/researchers/compliance/animal/sop/
Personal cleanliness is an important barrier to infection and washing of hands after handling any animal will reduce the risk of disease spread and self-infection. All employees working with animals, as well as visitors to the facility, should wear protective clothing, minimally a lab coat. By the way, no visitor is allowed unless approval is given by the chair of the UACC. Contact McGill University’s Media Relations Office for details ( http://www.mcgill.ca/newsroom/contacts/ ).
All contaminated material must be decontaminated before disposal. Necropsy of animals infected with highly infectious agents should be carried out in certified and tested biological safety cabinets. Necropsy material for disposal should be sealed in plastic bags, properly labeled and incinerated. The necropsy room should be properly equipped to provide adequate refrigeration and hand-washing facilities.
PHYSICAL INJURIES & CHEMICAL HAZARDS – PRECAUTIONS, REPORTING & DISPOSAL
Physical injuries related to the handling of animals may be kept to a minimum by ensuring that:
- everyone is trained and experienced in handling the species with which he works, and that he knows the particular hazards associated with each species;
- everyone is familiar with the hazards of the experiment, and is provided with (and use) a proper working area, protective clothing and equipment;
- a mechanism is in place in every unit to deal with animal-inflicted injury, and for referral for any further medical treatment if this is required.
Responsibility for ensuring that first aid kit(s) are available and always properly stocked must be clearly identified. The location of the first aid kit(s) should be prominently marked and all personnel using the facility should be made aware of these locations.
Injuries from chemicals can be avoided by treating all chemicals with care, by knowing their properties and adhering to the accepted safety practices for handling that type of product. WHMIS (Workplace Hazardous Material Information System), legislative and institutional requirements must be met.
Care should always be taken in handling such common chemicals as industrial detergents used in cage washers, cleaning agents, and powerful disinfectants. These substances should be stored separate from animal feed and bedding materials. Volatile liquids used as anesthetics or for euthanasia, and other toxic and volatile materials, should be stored in well-ventilated fume hoods or cabinets designed for that purpose.
Within 24 hours of an incident, IT IS VERY IMPORTANT TO COMPLETE AN ACCIDENT REPORT FORM. For McGill,tThis form can be downloaded from the Environmental Safety Office’s Web site at www.mcgill.ca/ehs/. This 2 page form will document the incident (in case the information is needed later) and measures can be take to avoid recurrence.
Disposal of animal carcasses entire or not, is done in a specific way. Containers, site of disposal, day and time of pick up, labeling, inclusion of other hazards (such as radiation) are important to know. The SOP for your animal facility and the protocol of the project should be carefully read and procedures followed. Any deviance must receive approval beforehand.
OCCUPATIONAL HEALTH PROGRAM FOR ANIMAL RELATED ACTIVITIES
Activities involving the care and use of animals in research and teaching pose particular health risks not normally encountered in other activities. The magnitude of risk is dependent on:
- the nature of contact (direct or indirect) with animals, their tissues, excreta, body fluids, hair, animal cages, dander and
- the species involved.
In recognition of its responsibility to provide a safe working environment, McGill University has adopted this policy for protection of faculty, staff, and students from health risks which may result from working with animals or working in animal care activities. Please see the Occupational Health Program website at www.mcgill.ca/ehs/
- Protection of individuals from hazards associated with animal care and use.
- Ensuring that individuals are thoroughly informed of the risks associated with their work.
- Provision of preventative medical services and delivery of prompt and adequate medical care and advice.
- Rehabilitation of employees whose health has been compromised due to occupational exposures.
- Ensuring that individuals are physically able to perform their assigned tasks without undue risk to themselves or fellow workers.
- Protection of animals from diseases they may contract from humans.
General Statement of Policy
Individuals involved in animal care and use are given protection from health risks through the following mechanisms:
- education and training,
- isolation and containment of high risk activities,
- evaluation of health status prior to exposure,
- periodic health assessment,
- provision of emergency health care,
- administration of immunization or other prophylaxis for protection from specific risks,
- exclusion of unauthorized persons from animal facilities,
- exclusion of persons with active infectious diseases that are hazardous to fellow workers or to animals,
- use of appropriate apparel, equipment, and facilities,
- veterinary management of animal health, and
- maintenance of records of illnesses, occupational diseases and injuries.
This policy requires confidentiality in the handling of medical information. Individuals required to undergo any medical examinations, treatment, or monitoring must be fully informed of the associated benefits and risks.
The services are to be provided at no cost to the individual. All activities related to this program are to be available during normal working hours.
Education and Training
Persons working with animals or in animal facilities should be fully informed as to the nature of possible risks associated with proposed duties. It is the responsibility of each laboratory director to inform research personnel of the specific risks involved and the applicable procedures.
Training in the principles of radiation and chemical safety is the responsibility of the local Environmental Safety Office.
The Biohazards Committee is mandated to develop protocols for research activities involving biohazards.
The University Animal Care Committee is responsible for verifying that those conducting research with animals are duly qualified.
Medical Care Elements of Program
This section is applicable to routine health surveillance activities involving animal care and use. Animal bites or other accidents involving animals are not covered in this section and should be considered for emergency treatment.
TABLE: Guideline for vaccination per species and level of contact:
1. Pre-placement assessment: medical history questionnaire and (if clinically indicated), medical examination.
2. Tetanus immunization (if not already up to date). Booster every ten years.
3. Selective pre-placement rabies immunization. Repeated as required.
4. Pre-placement PPD skin testing (2-step).
5. Hepatitis A vaccination; booster at 1 year follow-up
6. Q fever immunization
"Direct Contact" refers to those handling live animals, unpreserved tissues or body fluids, animal cages, cage accessories, animal waste or carcasses.
"Indirect Contact" refers to those who work in areas where animals are used or housed. These people are potentially exposed by means of accidental contact or aerosols.
N.B. This table is presently undergoing review
IF YOU ARE WORKING IN A HOSPITAL OR HOSPITAL RESEARCH INSTITUTE, please contact your local Health Office.
McGILL UNIVERSITY NON-HOSPITAL-RESEARCH-INSTITUTE members may apply to the Occupational Health Program for Animal Related Activities by going to the Environmental Health and Safety website: http://www.mcgill.ca/ehs/forms/forms/frequentlyused/
PERSONAL HYGIENE AND PROTECTION
Persons working with animals are required to maintain a high standard for personal cleanliness to reduce the risk of contracting diseases transmitted by animals. It is essential that facilities and supplies for meeting this obligation be provided. Clothing suitable for use in an animal facility are to be worn by all persons coming into contact with animals. For animal care staff, the clothing should be separate from that worn outside the animal facility and should be supplied and laundered by the institution. Clothing exposed to potentially hazardous microbial agents or toxic substances is to be decontaminated prior to leaving the premises for laundering. Disposable gear, such as gloves, masks, head covers, coats, coveralls, and shoe covers should be used where appropriate. Hands should be routinely washed after handling animals or cage accessories to reduce the risk of disease transmission.
Clothing should be changed as often as is necessary to maintain personal hygiene. Outer garments worn in animal rooms should not be worn outside the animal facility. Washing and showering facilities appropriate to the program are to be made available.
Provision of advice on specific procedures to be followed is the responsibility of the unit director or principal investigator.
Eating, drinking, smoking, or application of cosmetics in animal rooms are not permitted.
Infections that are secondarily transmitted from animals to humans are referred to as zoonoses and can seriously affect personnel and their research.
While most infectious agents show a considerable degree of species specificity, they also may, from time to time, vary widely in virulence and in their capacity to break through species barriers. Thus, infections that have not commonly been considered to be zoonotic hazards may sporadically affect susceptible persons or animals. Persons potentially at higher risk are those who suffer from defective immune systems and those who are under severe stress or who have non-overt clinical disease. Numerous pathogenic microorganisms, such as those responsible for tuberculosis, brucellosis, rabies, etc., which are normally perpetuated by direct transmission from one or more species of vertebrate animals, are also readily transmissible to humans.
Transmission of infections from animals to humans can generally be avoided through proper veterinary care and adherence to SOPs for control of transmission. However, when animals are obtained from areas in which zoonotic diseases are known to exist, e.g., in NHP (Non-Human Primate) acquired from the wild, special attention is required. Work involving exposure to hazardous microorganisms might require prior immunization of the staff, if a vaccine is available.
Caution should be exercised in assigning women of childbearing status to animal care duties that might expose them to potential or known teratogens. For example Toxoplasma gondii, a protozoan that infects most species of warm-blooded animals, including humans, is spread primarily by oocysts shed in cat feces. These oocysts sporulate in two to four days and may survive for more than a year. Human toxoplasmosis can result in spontaneous abortion, prematurity, stillbirth or congenital defects.
The life cycle of the causative organisms implicated in a number of indirect zoonoses may involve transmission through one or more other vertebrate and/or invertebrate intermediate hosts before affecting humans (for example, in taeniasis, tularemia, and vesicular stomatitis). Amongst invertebrate vectors of zoonotic disease, the biting insects are the main offenders. A list of some of the diseases transmitted to humans from animals can be found in section "12- Biological Hazards of Working with Experimental Animals " http://www.ccac.ca/en_/training/niaut/stream/cs-ohs.
The role of cold-blooded vertebrates in the epidemiology of zoonoses should not be overlooked. In particular, turtles infected with salmonella may constitute a human health hazard in the student laboratory as well as in the animal facility.
Allergies to laboratory animals are a significant occupational health concern for people regularly working with the common laboratory animal species. Laboratory animal allergy (LAA) is an immediate-type hypersensitivity reaction, IgE-mediated, which develops upon exposure to a laboratory animal, its fur or dander, its urine, saliva, serum or other body tissues. Typical symptoms range from mild (e.g., upper respiratory signs such as sneezing, itchy and/or runny nose and eyes, and skin reactions such as red, raised and itchy wheals after contact with animals, their tissues or their excreta), to severe [e.g., wheezing, shortness of breath, and a feeling of chest tightness (asthma)]. Persons experiencing such symptoms should be advised to contact their physician for diagnosis and treatment.
Measures which can reduce the degree of exposure to laboratory animal allergens include:
- use of protective gear such as gloves, face masks, gowns, shoe covers, etc., worn only in animal rooms;
- regular hand-washing, and showering after work;
- use of improved filtration in animal room ventilation systems, and the use of special filtered caging systems; and
- educational programs for employees identifying high risk (e.g., high allergen load) areas and tasks, and strict use of preventive measures.
Concerns related to lab animal allergies can be brought to the attention of the physician in the Occupational Health Program at the Environmental Health and Safety Office, if you have joined, or your personnel physician. As noted above, identifying high risk areas and tasks, and the use of SOPs in these areas, along with education, are useful in reducing the severity of the problem.
An animal_allergy_poster should be in evidence in your facility to remind you of the precautions.
Good Research Practices
It is very important that all research animals be monitored on a regular basis. While at least cursory observation of the animals is made by the animal attendants during cage cleaning procedures, this type of inspection may not detect problems in the early stages.
At a minimum, one of the researchers responsible for the animals should check each and every animal once per week.
Depending on the nature of the research being conducted on the animal more frequent monitoring may be necessary, for example: post-operatively, when using strains of animals that are known to develop specific problems at a given age, or when working with colonies of aged animals. If more frequent monitoring is required, the minimum interval of inspection may be determined by checking the “Clinical Endpoints” section of the protocol the animals are covered by. It is important that responsibility for this task be clearly defined and, when that person will be away for any length of time, another person designated to take over. Symptoms of an animal that is feeling pain or stress may vary by species. Some symptoms of pain include isolation from others in the same housing unit, abnormal body posture (i.e. hunching or stiffness), and vocalization, especially if disturbed. If housed singly, it may be possible to detect decreased appetite or reduction in fecal or urine output. Piloerection or lack of grooming is often a sign that animals that are not feeling well. The following table shows some of the common signs of pain and stress and gives an expectation of their occurrence. This table is not intended to be a definitive guide to determining the presence of pain and/or stress, especially since different species may exhibit different responses. Additional information on the species specific signs of pain is available from the CCAC at http://www.ccac.ca/en_/standards/guidelines/additional/vol2_mice.
WORKMAN, P., TWENTYMAN, P., BALKWILL, F., et al. (1998). United Kingdom Coordinating Committee on Cancer Research (UKCCCR) Guidelines for the welfare of animals in experimental neoplasia (Second Edition, July 1997). British Journal of Cancer 77:1-10.
Body Condition Score (BCS)
The McGill University Animal Care Committee (UACC) recommends the use of Body Condition Score (BCS) for clinical endpoints. Body condition scoring is a rapid, non-invasive and effective assessment of an animal’s physical well being.
In many instances, BCS is a better clinical endpoint than body weight. The use of body weight alone does not discriminate between body fat or muscle stores. Weight loss can be masked by abnormal (e.g. tumour growth, accumulation of ascetic fluid, organomegaly) or normal (e.g. pregnancy) weight gain. Alternatively, an animal that may have exceeded a 20%weight loss yet maintains a reasonable BCS will not necessarily require immediate euthanasia. Thus, BCS is a more comprehensive and accurate marker for animal health than a fixed percent of body weight loss.
A BCS less than 2 would usually be considered a clinical endpoint. Other clinical endpoints can also be reported such as decreased exploratory behavior, reluctance to move (decreased locomotion/mobility), pronounced hunched posture, piloerection (hair standing on end), moderate to severe dehydration (sunken eyes, prolonged skin tent, lethargy), unrelenting pain (e.g. distress vocalization). Endpoints specific to the project’s procedures should also be included.
Body Condition Score for Mice:
Body Condition Score for Rats:
FACTORS INFLUENCING RESEARCH RESULTS
- An ideal animal model is an individual that is sound both physically and mentally and is entirely suitable for the project
- Healthy laboratory animals will necessarily generate sound experimental data as opposed to their unwell or diseased counterparts
- Factors to consider in selecting an animal model:
- Accurate reflection of the system/organ in question
- Natural vs. induced disease model
- Genetic variables of model
- Species availability
- Size of animal model
- Life span of animal
- Housing costs
- Husbandry expertise
- Environmental enrichment
- Special requirements (diet, housing)
- Health status
- Viral, bacterial, parasitic, fungal pathogens
- Reflection of a single individual or for an entire animal colony
- Relevant for imported and exported animals
- Special importance in immunodeficient animals
- Species specific diseases
- Inbred vs. outbred
- Genetically modified animals
- e.g. disease susceptibility, tumour incidence, lifespan
- Nutritional plane
- Disease susceptibility generally increases with age
- Reproductive status (e.g. pregnancy, lactation, neutering)
- Body condition score
- Obesity / emaciation vs ideal body weight
- Acclimation period
- Adequate period of handling by technically skilled individuals prior to experimentation
- To minimize negative physiologic effects of transportation stress and introduction to a novel environment
- To allow for establishment of a baseline in terms of physiological parameters
- Relative humidity
- Disease susceptibility (ringtail in rats if RH less than 40%)
- Thermoneutral zone varies between species
- Conventional vs. barrier facility
- Static microisolators vs. ventilated racks
- Air quality
- Ammonia concentrations, allergens, pollutants
- Number of fresh air changes per hour
- Recommended 10-15 per hour
- Removes ammonia, heat, CO2 and airborne particles (allergens)
- Cage population density
- Negative impact on reproductive performance
- Often out of human auditory range
- Negative impact on reproductive performance
- Audiogenic seizures (strain dependent)
- Traffic flow patterns
- In animal facility and research laboratory
- Quality, quantity, availability
- Nutritional plane, availability, palatability
- Feed contaminants
- At cage or room level - Scientifically proven positive impact upon research animals
- Light / dark cycle
- Natural circadian rhythms
- Photoperiod (12 hours light: 12 hours dark) in rodent rooms
- As most rodents are nocturnal, consider this when planning the timing and duration of manipulations and experimental interventions
- Manipulations should be carried out at same time each day to ensure consistent results
- Intensity of light source (above 300 lux can cause retinal degeneration in albino animals)
- Light pollution (interference with tumour growth rates)
- Research community
- Familiarity with appropriate animal models, experimental design and objectives
- Knowledge of humane endpoints
- Competent research personnel trained to work with animals and carry out experimental procedures
- Post operative care and monitoring skills
- Animal health team (veterinarians and veterinary technicians)
- Competent and humane animal handling
- Familiarity with species specific behaviours and diseases
- Knowledge and recognition of illnesses and diseases
- Therapeutic intervention and/or alleviation of clinical signs
- Post operative care and monitoring skills
- Technical expertise in animal handling and biomethodology techniques (blood collection, compound administration etc)
On a larger scale, the institutional sentinel program, micro monitoring procedures and quarantine policy will all have an impact upon the health of both individual animals, and the animal community as a whole.
Infectious disease in a laboratory animal facility can kill many of the animals, alter the animal's immune system interfering with research results, can spread to people and there may be a need to depopulate the facility in order to eliminate the threat.
It is important to be proactive in reducing stress in research animals. When preparing a research protocol, minimally invasive techniques should be considered where possible. Pilot studies may be used to find the best method of performing an experimental procedure before starting studies on larger group.
When animals are first received in the animal facility, a period of acclimation is necessary to allow the animals to recover from transport stress and to adjust to their new surroundings and, possibly, new cage mates. The table above shows the recommended acclimation periods for various species. Note that basic handling may be begun inside the acclimation period for most species. Research procedures should wait until after the acclimation period.
Maintaining consistency of handlers is a very good way to reduce stress among research animals. Animals often become accustomed to a particular handler and become less stressed when always handled by that person. Personnel should have assigned duties and minimize asking others to take over for them when at all possible. Consistency in the way techniques are performed on the animals is also very important. Husbandry procedures are less stressful if performed regularly by the same person. All handlers should work in a calm and quiet manner, avoiding unnecessary noise and a busy, ‘bustling’ work attitude.
Pair or group housing of social animals is an important method of reducing stress. If removing an animal from a housing unit for testing, bringing a cage mate along can reduce the stress levels felt by the experimental animal.
Using acclimation and operant conditioning to introduce an animal to an experimental procedure can significantly reduce stress levels. Before starting potentially stressful procedures, animals should be given the opportunity to get used to the apparatus on several occasions beforehand, especially in the company of an animal which is used to that apparatus. Placing experimental apparatus in the housing area, at a gradually decreasing distance, is one possibility. Animals, especially larger animals, can be trained in gradual steps to perform required tasks, such as presenting a limb for injection or remaining quietly in a sling or other restraint for prolonged periods of time. This type of acclimation must only start after the initial acclimation upon arrival is completed.
Experimental animals were traditionally kept in caging which provided little or no social or physical stimulation. The use of such caging was justified on the basis of reduction of disease spread, ease of sanitation, prevention of fights between animals, easy recognition of illness through measuring food and water intake, etc. However, at the time, little consideration was given to the behavioural and psychological well-being or the stress induced by social isolation and physical deprivation. It is recognized now, that the well-being of animals is greatly improved if they are provided with opportunities for interaction with each other and their environment. Furthermore, there is an increasing volume of literature denoting the deleterious effects of impoverished environments on experimental results.
Although the term "environmental enrichment" is used to describe efforts aimed at improving the living conditions for animals, the move is really from a very impoverished environment to a less impoverished environment. It is unlikely that the level of complexities encountered by wild counterparts will ever be achieved within the laboratory. Furthermore, it is possible that the well-being of an animal will not be increased by our ideas of increased complexity in its environment. Nevertheless, the wild species are often taken as the norm against which the environment of the captive animal is measured. Some argue that the wild and laboratory animals are no longer the same behaviourally, but most wild behaviours are seen in the laboratory animal.
The presence of a normal range of behaviours and the absence of abnormal behaviours or stereotypies (behaviors that do not appear to be goal-directed) is a reasonable indication that the animal is coping with its environment. To make such judgements, we must be able to recognize normal and abnormal behaviours. Those species that are prey animals in nature, seldom reveal that they are hurting in any way as this would be an invitation for predation.
Another approach to evaluating well-being is to use the Five Freedoms of the UK Farm Animal Welfare Council. These freedoms were defined to give guidance to farmers on the goals of husbandry. However, the freedoms are easily adapted to other animals and have been accepted by various groups including the World Veterinary Association and Humane Societies.
The five freedoms are:
1 - Freedom from hunger and thirst (by ready access to fresh water and a diet to maintain full health and vigour)
2 - Freedom from discomfort (by providing an appropriate environment including shelter and a comfortable resting area)
3 - Freedom from pain, injury and disease (by prevention or rapid diagnosis and treatment)
4 - Freedom to express normal behaviour (by providing sufficient space, proper facilities and company of the animal's own kind)
5 - Freedom from fear and distress (by ensuring conditions and treatment which avoid mental suffering)
The freedoms are general enough to allow them to be used for any animal species and to allow for interpretation related to particular species. They must be applied carefully with an understanding of the biology of each species and care must be taken to avoid using our own ideas as animal standards. For example, it is customary to keep newly weaned piglets at a temperature of about 27° 0 C, day and night. However, when the piglets had the opportunity to control the temperature themselves, they preferred a temperature of about 29°0 C during the day and about 15°0 C at night.
Terms like "discomfort" make us think about the animal's living conditions and while we tend to think of the extremes of heat and cold, or wet and dry, there are grades of discomfort in between the extremes as we know from our own experience. A cool room is uncomfortable if we do not wear enough clothes and a hairless animal without any means of building a nest or others to huddle with may be uncomfortable at the normally recommended temperatures in the animal facility.
We can assume, given our present knowledge, that the health, nutrition and general environment needs of the common laboratory animal species are being met in present day laboratory animal facilities. The major challenge for us is to provide them with social and physical opportunities to live and behave in a normal manner. To do that we must have some knowledge of what a particular animal needs based on understanding their preferences. Most important, all animals require social interactions although for some this interaction is intermittent and occurs only at breeding times. Most wild animals occupy their days in the search for food and water. The threat of predation is a fact of life for many small animals, including those in the laboratory where we are the predators. To be frightened without having any means of protecting yourself is a stressful experience. Lack of space or structure to exercise or play, in the case of young animals, is detrimental to bone and muscle development and maintenance.
The major factors to be considered then are:
- Opportunities to socialize or not
- Opportunities to occupy time during waking hours
- Opportunities to hide
- Opportunities and structure for exercise
Improving the environment is not just a nicety for research animals. There is a considerable body of literature now that demonstrates the influence of an animal's physical and social environment on research results. One of the earlier demonstrations showed that social and physical stimulation of rats resulted in a thicker cerebral cortex with more dendritic connections. Tumours in isolated mice grow faster than the same tumours in mice housed at appropriate densities. Isolation of mice has been shown to increase the toxic effects of some drugs.
It has also been shown that environmental enrichment is beneficial at any stage of an animal's life. The effects may be different between young and old animals but the old will also benefit. For this reason it is important to consider environmental enrichment as a variable in an experiment and to account for it. It is not an option, however, to omit environmental enrichment to reduce the variables in a study unless the investigator is prepared to include all the deleterious effects of an impoverished environment on the study. Even then, it would be difficult to say that the results represent the normal state of the animal. However, if an investigator feels that attempts at environmental enrichment will jeopardize the results of a study, then this should be justified to the Animal Care Committee.
Environmental enrichment encompasses more than just the physical and social environment of a group of animals. Because we interact with them at various levels, we may have a profound effect on their life. We should treat animals in a manner that minimizes any discomfort or stress they may experience at our hands. All the environmental enrichment in the world will not be of any value if an animal fears the arrival of a human being at its cage. It may not be just the presence of a person but it may also include the sounds and smells associated with an experimental procedure, for example.
Our activities in the animal facility may be disturbing, even if they do not directly involve the animals. Noise is disturbing to animals and we should minimize extraneous noises as much as possible. Some people bustle and have an air of urgency about them that is unsettling to animals. Doors are allowed to slam shut or objects fall on the floor. Equipment like cage washers, vacuums etc. may be upsetting, particularly to pregnant animals.
If we try to see things as the animals might see them, we will probably be able to improve their living conditions. In return, people working with the animals, particularly the animal care technicians, feel better about their jobs when they see the animals responding to their enriched environment.
It must be emphasized that changing an animal's environment, whether it be giving it a clean cage devoid of familiar "homey" smells or adding toys or other objects for enrichment, will be a variable that should be accounted for. It is important, then, not to make changes to the environment without the agreement of the principal investigator and if changes are made, they should be applied consistently to all animals in the study. It should be remembered that there may be effects of withdrawing enrichment, for example, if the animals move from a facility with very complex environments to one where there is minimal complexity.
In the past, animals were kept in cages or pens that provided very little substrate for them to engage in many of their natural behaviours. Environmental enrichment is a phrase used to cover a wide range of additions or modifications of the environment to allow animals a more varied life. Although many of the changes are indeed to the physical environment, changes in social opportunities for the animals are also important; and social opportunities include interactions with people. A major benefit of environmental enrichment is the reduction in stress in the animals with beneficial influences on the research projects they are involved in.
Animal Users must know that environmental enrichment is part of the set of guidelines from the CCAC. It is not a matter of choice but a requirement. Animal facilities will include enrichment devices to single housed animals as part of the normal process. Investigators who do not want such devices, clear justification must be supplied to the local animal care committee.
Examples of enrichment
- Study design
- Housing - wire-bottom vs. solid bottom cages
- Social interaction - single vs. pair vs. group housing
- Feed - ad libitum
- Enrichment devices - tubes, running wheels, marbles
- Human interaction - minor value
- Study design
- Housing - wire-bottom vs. solid bottom cages
- Social interaction - single vs. pair vs. group housing
- Feed - ad libitum vs. diet optimization
- Enrichment devices - tubes, marbles, hanging toys
- Human interaction - possible value
- Study design
- Housing - flooring considerations
- Social interaction - single housing with visual contact
- Feed - limit feed, treats
- Enrichment devices - toys, music
- Human interaction - probable benefit
For other species, a search on the Web will bring plenty of information regarding what is best for the animal you are working with.
A controlled substance is any type of drug that the federal government has categorized as having a higher-than-average potential for abuse or addiction and has taken measures to control its use such as limiting the type of users and requiring a written application for permission to use.
Due to the potential for abuse, permits and/or valid practice licenses are required for the purchase, distribution and administration of opioids as well other controlled substances such as ketamine. Each investigator who plans to use a controlled substance must complete an “Application Form for an Exemption to Use a Controlled Substance for Scientific Purposes” and submit it to the Office of Controlled Substances at Health Canada. Justification for the amount of drug to be purchased must be provided as well as the name and address of the supplier (e.g. The Animal Resources Centre is a licensed distributor for many of the controlled drugs used in research at McGill University). Both applicant and supplier will receive copies of the permit once the request has been approved. Call Health Canada at 613-952-2219 for further information.
Up-to-date record keeping logs are mandatory. A sample log is available online at: Controlled drugs dispensing form [.xls] or see sample form below. The log should permit recording of the type of drug the record is for, the name of the Principal Investigator under whose permit the drug was ordered and the date of issue/receipt.
A. FACILITY SPECIFIC ISSUES
A laboratory animal facility must facilitate research by minimizing undesirable experimental variables while providing for the physiological, social and behavioural requirements of the animal. Different research projects and/or different species of animals often require differing facilities and environments. To accommodate such needs, an animal facility must have separate areas for carrying out different functions, specialized rooms and equipment, and closely controlled environments.
Animal facilities providing the appropriate environment are expensive to build. It is, therefore, imperative that every effort be made to ensure that any proposed new facility is programmed, designed, and built to meet the size and scope of current animal use, and yet to be versatile enough to allow flexibility in the years to come.
A number of alternative design approaches to achieve any given functional need are available. It is strongly recommended that the CCAC be involved at an early stage in the planning phase and that plans be evaluated by the Council before the start of construction.
Important issues include:
- location (secured, clean and dirty material kept separate),
- mechanical services (heating, ventilation and air conditioning systems,
- design (size of room, easy to sanitize, easy to access for authorized personnel)
- major functional divisions (animal holding rooms separate from procedure rooms, reception area, one species per room, hazardous material used in separate rooms, quarantine rooms, washing facilities, waste disposal, food and bedding storage area, office area)
- security (controlled access to facility)
- construction (floor drains, no cracks in walls & ceilings, windows that do not open)
- caging (appropriate for species, easy to clean, allow view of animals, with a solid bottom)
Information on housing of large domestic animals and fowl may be found in Agriculture Canada's Canadian Farm Building Handbook (Agriculture Canada, 1988) as well as Social and Behavioural Requirements of Experimental Animals on the CCAC Web site at http://www.ccac.ca/en_/standards/guidelines.
Information on shipping crates and transport cages for a wide range of domestic, wild and laboratory animals may be obtained from the most recent volume of Live Animals Regulations (1992) of the International Air Transport Association (IATA).
B. THE ENVIRONMENT
There are many physical, chemical, and biological factors which may influence experimental animals and thus modify the results of the investigations. The experimental results obtained are, in principle, only valid for the conditions under which they were obtained and only useful for comparison if all the relevant information concerning experimental conditions is made available.
Among the environmental factors which should be recorded for possible inclusion in scientific reports are: temperature (C and range), relative humidity (% and range) and whether or not these are regulated; air exchanges/hour, proportion of fresh and recirculated air, and gas or particle concentrations in the air; lighting (natural and/or artificial, photoperiod, and intensity); water type, quality, and pretreatment; bedding type, quality, and pretreatment; housing density; housing equipment; and physical measures to protect microbiological status. The microbiological status of the animal should be reported [conventional, Specific Pathogen Free (SPF) for stated pathogens, or gnotobiotic with microorganisms specified].
Environmental requirements vary with the species and the experimental protocol. Environmental parameters are usually measured at the level of the room. More important, however, is the microenvironment established at the cage level, since the conditions between the two may differ dramatically. A summary of some environmental parameters for individual species is given in Appendix I of the CCAC guidelines.
The design of the animal facility should permit adjustment of environmental controls to meet the needs of the species and the experimental protocol. Ideally, each animal room would be controlled independently. In facilities not originally constructed with this capability, this ideal could be approached through proper management and the installation of ancillary automatic light timers, rheostats, thermostatically controlled exhaust fans, humidifiers, and air conditioning units.
- population density and space limitations
DESIRABLE CRITERIA FOR RODENT CONTACT BEDDING (Kraft, 1980) : Moisture absorbent, dust free, unable to support bacterial growth, inedible, non-staining, non-traumatic, ammonia binding, sterilizable, deleterious products not formed as a result of sterilization, easily stored, non-desiccating to the animal, uncontaminated, non-nutritious, non-palatable, unlikely to be chewed or mouthed, non-toxic, non-malodorous, nestable, disposable by incineration, readily available, relatively inexpensive, fire resistant, remains chemically stable during use, manifests batch to batch uniformity, optimizes normal animal behaviour, non-deleterious to cage-washers, non-injurious and non-hazardous to personnel
Unsterilized materials are a possible source for the introduction of disease into rodent colonies. Wild rodents enjoy nesting in packages of bedding, and cats will defecate in loose bedding.
The effects that microbiological agents can have on experimental results and the health of laboratory animals have been widely documented. Control of the microbiological status of the experimental animal and its environment is necessary for valid scientific results and animal well-being. The sources of microbial contamination include vermin, experimentally infected and spontaneously ill laboratory animals or their tissues or tumours, air, food, water, bedding, ancillary equipment, and personnel. Good facility management practices and constant surveillance are necessary to minimize the introduction of unwanted microbes. Insect and rodent vermin should be strictly controlled or excluded from the facility.
Whenever possible, the health status of all animals should be ascertained before the animal is brought into the facility. Animals having an unknown health status should be quarantined and tested before being admitted to the facility. Additionally, all tumour and cell lines should be tested before being introduced. Research on contagious diseases must be carried out in appropriate containment facilities.
Containment is required for animals exposed to known infectious microorganisms. Required containment and management procedures vary with the biohazard classification of the microorganism, based on the degree of risk to man and other animals. Personnel may be required to shower before leaving the containment unit. All cages and materials are sterilized upon leaving the area. Air pressures are balanced so that the highest pressure is outside the containment area. Air exiting the facility is diluted with clean air, filtered, or incinerated.
Because it is hazardous to staff and animals, UV light is not generally recommended for routine disinfection of laboratory air. The infectious disease unit should be segregated as much as possible from the rest of the animal facility. Specific requirements will differ with the degree of risk. Depending on the hazard, containment of small groups of animals may be accomplished with flexible film isolators or microisolation cages. The use of laminar airflow racks to prevent cross-contamination between cages should be carefully evaluated as the transfer of certain pathogens may be enhanced in some instances. Infectious disease units should be disinfected immediately following use.
Recommendations for control of biohazards can be found in Laboratory Biosafety Guidelines (http://www.hc-sc.gc.ca/pphb-dgspsp/publicat/lbg-ldmbl-96/index.html) and elsewhere. Biological safety cabinets approved for the appropriate biohazard level must be used for experimental manipulations. These cabinets must be inspected and tested annually by trained personnel.
Persons working in infectious disease units should be protected with a comprehensive occupational health and safety program.
CHEMICAL AND RADIOISOTOPE UNITS
In Canada, laboratory use of radioisotopes is regulated by the (federal) Canadian Nuclear Safety Commission (CNSC, http://www.nuclearsafety.gc.ca/eng). The CNSC issues licences to the institution for the possession of radioactive material. When radioisotopes are used in animals experimentally, Standard Operating Procedures (SOPs) to ensure that related hazards are minimized should be defined and enforced; these SOPs are considered by the CNSC when it issues the Radiation Licence. As well, the CNSC recommends that the institution's Radiation Safety Officer sit on the Occupational Health and Safety Committee in an ex-officio capacity.
The Workplace Hazardous Materials Information System (WHMIS) is regulated by federal and provincial health and safety authorities. It legislates labeling requirements, availability of Material Safety Data Sheets (MSDS), and training programs required for personnel to work safely with certain hazardous materials.
The chemical and radiation hazard area should be separated from other animal housing and work areas. The hazardous area must be clearly posted and entry restricted to necessary personnel. Contaminated cages should not be transported through corridors. Safe transport equipment and procedures should be developed if necessary. Laminar flow cage-changing stations are recommended to protect the staff from aerosolized contaminants.
Category of Invasiveness
Canadian Council on Animal Care:
- Established in 1968, the CCAC has established enhanced animal care and use programs within the scientific community through its voluntary compliance, education and code of ethics
- The CCAC is a national peer review organization with a mandate to work for the improvement of animal care and use on a Canada wide basis
- Its Assessment Program focuses on institutional animal care and use programs, and the functioning of the local Animal Care Committee
All animal protocols using vertebrates or invertebrates must be assigned a CCAC category of invasiveness in animal experimentation
CCAC Categories of Invasiveness:
- Category A
- Category B
- Category C
- Category D
- Category E
Experiments on most invertebrates or on live isolates.
- Tissue culture
- Tissues obtained at necropsy or from the slaughterhouse
- Use of eggs, single celled organisms, protozoa and metazoa
Experiments that cause little or no stress or discomfort.
- Maintenance of domestic herds/flocks
- Commercial herd/flock production
- Short term and skillful restraint of animals for observation or physical examination
- Blood sampling
- Injection of material via intravenous, intramuscular, subcutaneous, intraperitoneal and oral routes with no known adverse reaction
- Short periods of water and/or food deprivation equivalent to periods of abstinence in nature
- Acute non survival studies
Experiments which cause minor stress or pain of short duration.
There should be no significant alterations to the animal’s appearance, or physiological parameters such as respiratory and heart rates, food and water intake and urine and fecal output.
In addition, category “C” animals must not demonstrate any self mutilation, dehydration, anorexia or alterations in behaviour (aggression, withdrawal, increased or decreased recumbency or dormancy, social interactions)
- Cannulation or catheterization of blood vessels or body cavities under anesthesia
- Minor surgical procedures under anesthesia (biopsies, laparoscopy)
- Short periods of restraint beyond that for simple observation with minimal distress
- Short periods of water and/or food deprivation exceeding periods of abstinence in nature
- Exposure to non lethal doses of drugs or chemicals
Experiments which cause moderate to severe distress and discomfort.
Procedures in Category D should not cause prolonged or severe distress evident by the following clinical signs: abnormalities in behaviour, absence of grooming, dehydration, abnormal vocalization, immobility, prolonged anorexia, indications of severe local or systemic infection or extreme lethargy.
- Major survival surgical procedures under general anesthesia
- Behavioural stresses (maternal deprivation, aggression, predator-prey interactions)
- Physical restraint for prolonged periods (several hours or more)
- Exposure to chemical/drugs that impair physiological systems
- Production of radiation sickness
- Use of Freund’s Complete Adjuvant
- Induction of anatomical or physiological abnormalities that cause pain and distress
- Exposure to noxious, inescapable stimuli
- Procedures causing severe, persistent or irreversible disruption/malfunction of the sensorimotor centres
Experiments that cause severe pain near, at or above the pain tolerance threshold of conscious, unanesthetized animals.
E level protocols are NOT permitted at McGill
This category is not restricted to surgical procedures, and includes exposure to noxious chemicals or agents with unknown or ill-defined effects, or that markedly impair physiological systems causing extreme distress, pain and death. Death is often the endpoint.
- Use of muscle relaxants or paralytic agents without anesthesia
- Euthanasia method not approved by the CCAC
- Injection of noxious substances
- Induction of severe stress or shock
- Extremely invasive surgical procedures
- New biomedical experiments with a high degree of invasiveness
- Burn or trauma infliction on unanesthetized animals
- Any experiment where the pain cannot be relieved by analgesia
- A survival procedure with direct visual access to a major body cavity (cranium, thorax, spinal canal, abdomen, pelvis)
- Exposure of major vascular, lymphatic, muscular, skeletal, or glandular structures
- Removal and /or alteration to a functionally significant amount of viable tissue
- A surgical procedure that does not result in removal or alteration to a functionally significant amount of viable tissue
- Often no clear delineation between “major” and “minor” surgery
Both “major and minor surgery” definitions are used as adjuncts to CCAC Categories of Invasiveness.
A gentle death that is regarded as a act of humane killing with the minimum of pain, fear and distress.
- Unconsciousness and death are achieved rapidly
- Minimal restraint and psychological stress
- Simple, reliable, reproducible and irreversible
- Safe for the operator
"In experiments involving animals, any actual or potential pain, distress or discomfort should be minimized or alleviated by choosing the earliest endpoint that is compatible with scientific objectives of the research. "
- CCAC, 1998
The clinical endpoint lists the conditions, complications and criteria that would lead to euthanasia of an animal before the expected completion of the experiment, e.g. more than 20% weight loss, maximum tumour size, vocalizing, lack of grooming.
The experimental endpoint, is the estimated survival time for the animals.
Reasons for Euthanasia:
- Levels of pain, distress and suffering exceed acceptable limits
- Specific guidelines for physiological parameters (weight loss, reduced mobility, alterations in core body temperature etc)
- Specific guidelines for certain areas of research (neoplasia, toxicology)
- Experimental objectives / results are achieved
- Tissue harvesting
- No longer suitable for breeding
- Unwanted genotype / phenotype
- Questionable / undesirable health status
Recognition and Confirmation of Death:
- Cessation of heartbeat and respiration
- Absence of all reflexes
- Central, fixed dilated pupil (easier to detect in larger species)
- Chemical and physical methods
Acceptable Methods of Euthanasia:
- Species dependent
- Methods differ for anaesthetized vs. unanaesthetized animals
- Physical methods must cause immediate loss of consciousness through physical trauma to the brain
Recommended Methods of Euthanasia:
What best describes a good method of euthanasia is that it consistently produces a humane death.
- For chemical euthanasia, the intravenous route is always preferable to intraperitoneal route due to rapid induction and a quick, humane death.
- For physical euthanasia, prior anaesthesia is mandatory. If there is no anaesthesia administered prior to a physical means of euthanasia, scientific justification must be provided, with subsequent approval by the local Animal Care Committee.
Recommended Methods of Euthanasia (by species):
- Tricaine methane sulfonate (MS 222) immersion overdose. Neutralized to pH 7.5 with bicarbonate
- Benzocaine immersion overdose. Neutralized to pH 7.5 with bicarbonate
- Concussion followed by exsanguination, removal of the heart or cervical dislocation
- MS 222 immersion overdose. Neutralized to pH 7.5 with bicarbonate
- Benzocaine immersion overdose. Neutralized to pH 7.5 with bicarbonate
- Sodium pentobarbital overdose (intravenous, IV or intraperitoneal, IP)
- Concussion followed by pithing
- Decapitation and pithing
- Sodium pentobarbital overdose (IV or IP)
- Captive bolt followed by destruction of brain
Note: For amphibian and reptilian species, inhalational anaesthetics are an unacceptable means of euthanasia due to breath holding capacity, resulting in prolonged induction times.
- CO2 (in chicks up to 72 hours old)
- Volatile inhalational anaesthetic overdose
- Sodium pentobarbital overdose (IV or IP)
- Concussion followed by exsanguination or decapitation
- Anaesthetized birds:
- Potassium chloride
Note: exsanguination is unacceptable in birds due to rapid blood clotting
- Sodium pentobarbital overdose (IV or IP)
- Inhalation anaesthetic overdose
- Isofluorane followed by CO2 inhalation (animals more than 10 days old)
- Microwave irradiation in specially designed units
- Cervical dislocation
- Rapid freezing
- Sodium pentobarbital overdose (IV or IP)
- Inhalation anaesthetic overdose – prior sedation mandatory
- Potassium chloride
CARNIVORES: Dogs, cats, ferrets
- Sodium pentobarbital overdose (IV only) – prior sedation mandatory
- Inhalational anaesthetic overdose – prior sedation mandatory
Physical methods (field conditions only)
- Potassium chloride
LARGE MAMMALS: (Pigs, sheep, goats, cattle)
- Sodium pentobarbital overdose (IV only) – prior sedation mandatory
- Inhalation anaesthetic overdose (lambs and kids only) – prior sedation mandatory
Physical methods (field conditions only)
- Captive bolt followed by exsanguination
- Free bullet humane killers
Anaesthetized large mammals
- Potassium chloride
- · Sodium pentobarbital overdose (IV only) – prior sedation mandatory
Unacceptable Methods of Euthanasia for All Species:
- Air embolism
- Burning (chemical or thermal)
- Carbon monoxide
- Chloral hydrate
- Decompression / vacuum
- Diethyl ether
- Exsanguination without anaesthesia
- Household products and solvents
- Hypothermia / hyperthermia
- Magnesium sulfate
- Neuromuscular blocking agents
- Nitrous oxide
- Rapid freezing without anaesthesia
- Removal from water (gilled vertebrates)
End of the BASIC LEVEL theory course material.
Chick here to go onto the ADVANCED LEVEL of the theory course if you need it.
For information about practical courses (workshops), visit the section on the TRAINING PROGRAM/WORKSHOPS.